The ability of fresh isolates of B. gingivalis to establish abscesses in the mouse model was studied by comparing them with established laboratory strains of B. gingivalis. Eight fresh isolates obtained from plaque associated with periodontal disease and grown under similar conditions as established strains were injected subcutaneously on the back of the mouse. All of these strains produced secondary lesions on the abdomen. Septicemia was associated with seven of the strains. Two commonly used laboratory strains, W50 and W83, also produced secondary lesions and septicemia. Five other laboratory strains produced only localized abscesses. On histologic examination, the strains that produced disseminated disease showed invasion of connective disease by individual bacteria that were not in clumps. The strains that produced localized abscesses were characterized by growing in colonies or clumps in the abscess cavity. Four synthetic enzyme substrates were examined to determine whether the differences between invasive and non-invasive strains were due to differences in proteolytic enzyme production. No differences in enzyme production could be demonstrated with the selected substrates.
Five healthy children under 6 years of age, five healthy adults, and 10 adult periodontitis patients were examined for the prevalence and distribution of blackpigmented Bacteroides in the oral cavity. A total of 13 samples was obtained from each individual, including four supragingival and four subgingival dental plaques, dental occlusal surface, buccal mucosa, dorsal tongue, tonsil, and whole saliva. Black-pigmented Bacteroides were recovered from nine adult periodontitis patients. Healthy adults harbored the organisms in low incidence and proportions, whereas the children exhibited no cultivable black-pigmented Bacteroides. The organisms were isolated in highest proportions from dental plaque, especially subgingival plaque, and from the tonsil area, indicating that these sites constitute the organisms' primary ecological niche in the oral cavity. The predominant isolate was Bacteroides melaninogenicus subsp. intermedius followed by Bacteroides gingivalis and B. melaninogenicus subsp. melaninogenicus. B. melaninogenicus subsp. levii constituted low proportions of supragingival microflora of one adult periodontitis patient. A positive correlation was demonstrated between the proportion of black-pigmented Bacteroides (mainly B. melaninogenicus subsp. intermedius) and both the severity of gingival inflammation and the periodontal pocket depth, suggesting that these organisms may contribute to the pathogenesis of certain forms of periodontal disease.
The subgingival microflora and serum antibody response was examined in Periodontitis patients with noninsulin‐dependent diabetes mellitus (NIDDM), impaired glucose tolerance (IGT), and normal glucose tolerance (NGT). The predominant cultivable microflora was determined for subgingival plaque sampled from two deep periodontal pockets in each of eight adult Periodontitis patients with NIDDM. Indirect immunofluorescence for Bactewides intermedius, Bacteroides gingivalis, and Haemophilus actinomycetemcomitans was used to examine these same samples as well as 186 additional subgingival plaque samples from 47 patients with moderate to severe generalized Periodontitis including 25 subjects with NIDDM, six subjects with IGT, and 16 subjects with NGT. Serum antibody levels to 13 microorganisms including seven oral bacterial species and one nonoral control species were measured by enzyme‐linked immunosorbent assays (ELISA) in 377 subjects including 84 normal subjects without periodontal disease, 112 normal subjects with Periodontitis, 19 periodontally normal subjects with IGT, 65 Periodontitis patients with IGT, 15 periodontally normal subjects with NIDDM, and 82 Periodontitis patients with NIDDM. Three hundred eighty‐two bacterial isolates were recovered from the eight patients. B. intermedius was the most frequently isolated microorganism constituting 16% of the total isolates followed by Wolinella recta and B. gingivalis, which each accounted for 13% of the total. Streptococcus sanguis was the most prevalent microorganism, which was found in 75% of the sites. Subgingival plaque samples examined by immunofluorescence demonstrate a high prevalence of black‐pigmented Bacteroides and suggest that the proportion of B. gingivalis but not B. intermedius is higher in NIDDM with Periodontitis than in other groups. Serological studies of B. gingivalis from patients with NIDDM indicate that these isolates comprise a serogroup distinct from those found in adult Periodontitis patients without diabetes. Serum IgG antibody levels were significantly elevated in nondiabetic, IGT, and NIDDM patients with Periodontitis toward both B. gingivalis strains tested. Nondiabetic Periodontitis patients had significantly elevated levels of serum IgG toward B. intermedius strain 9336 while Periodontitis patients with IGT had significantly elevated antibody levels to B. intermedius strain 25261. Microbiologic and immunologic data from the present study suggests that B. intermedius. W. recta, and B. gingivalis are important in the etiology of Periodontitis in adult patients with NIDDM.
Two unique forms of periodontal disease, HIV‐gingivitis and HIV‐periodontitis, have been described in patients with Acquired Immunodeficiency Syndrome (AIDS). In order to determine the bacterial species associated with periodontitis in AIDS patients, the predominant cultivable microflora was examined in 21 subgingival plaque samples from 11 AIDS patients with periodontitis. The presence of putative periodontal pathogens including Actinobacillus actinomycetemcomitans, Bacteroides intermedias, Porphyromonas gingivalis (formerly B. gingivalis), and Wolinella recta was examined by immunofluorescence in 128 subgingival dental plaque samples from 50 AIDS patients including 32 patients with periodontitis. Of 666 bacterial strains isolated from the 21 subgingival plaque samples, Streptococcus sanguis II was the most frequently recovered species comprising 18.5% of the total number of isolates followed by Lactobacillus acidophilus (12.2%), Porphyromonas gingivalis (12%), Fusobacterium nucleatum (11.4%), Staphylococcus epidermidis (8.7%), Actinomyces naeslundii (7.5%), and Actinomyces viscosus (4.7%). Fusobacterium nucleatum was the most prevalent species and was found in 76% of the sites and 91% of the patients. Enteric species including Enterococcus avium and Enterococcus faecalis, Clostridium clostridiiforme and Clostridium diffide as well as Klebsiella pneumoniae also were recovered. Immunofluorescence assays detected similar carriage rates of A. actinomycetemcomitans, B. intermedius, and P. gingivalis in both gingivitis patients and periodontitis patients, while four times more periodontitis patients demonstrated W. recta. Subgingival yeast was a frequent finding in these AIDS patients, present in 62% of the subjects and 55% of the sites. This study indicates that subgingival plaque in AIDS patients with periodontitis can harbor high proportions of the same periodontal pathogens as are associated with periodontitis in non‐HIV infected subjects as well as high proportions of opportunistic pathogens. J Periodontal 1990;61:699–704.
Black-pigmented Bacteroides strains were grown on blood agar, and the colonies were evaluated for fluorescence from long-wave UV light. Most test strains of Bacteroides melaninogenicus subsp. intermedius exhibited a brilliant red fluorescence. B. melaninogenicus subsp. melaninogenicus fluoresced mostly red-orange. Bacteroides asaccharolyticus showed a yellow or red fluorescence. The intensity of the Bacteroides fluorescence weakened when the black pigment of the colonies developed. In contrast, neither young nor old colonies of the oral species Bacteroides gingivalis displayed fluorescence. Since B. gingivalis can produce severe oral infections and also can seed to nonoral sites, awareness of the inability of this organism to fluoresce is important for microbiologists utilizing UV light fluorescence to screen for black-pigmented Bacteroides spp. The present data also indicate that UV light fluorescence may be a rapid method of distinguishing some black-pigmented Bacteroides spp.
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